Treatment of HIV and acute myeloid leukemia by allogeneic CCR5-d32 - - PowerPoint PPT Presentation

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Treatment of HIV and acute myeloid leukemia by allogeneic CCR5-d32 - - PowerPoint PPT Presentation

Feb 02, 2023 291 likes •633 views

Treatment of HIV and acute myeloid leukemia by allogeneic CCR5-d32 blood stem cell transplantation Elena Knops Elena Knops, AREVIR, Cologne 2017 Background - the Berlin patient so far the only person presumed to be cured from HIV by

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Elena Knops, AREVIR, Cologne 2017

Treatment of HIV and acute myeloid leukemia by allogeneic CCR5-d32 blood stem cell transplantation

Elena Knops

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Elena Knops, AREVIR, Cologne 2017

Background

  • the Berlin patient – so far the only person presumed to be cured from HIV by

hematopoietic stem cell transplantation (HSCT) from a homozygous CCR5-d32 donor

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Elena Knops, AREVIR, Cologne 2017

Background

  • the Berlin patient – so far the only person presumed to be cured from HIV by

hematopoietic stem cell transplantation (HSCT) from a homozygous CCR5-d32 donor

  • failed attempts to reproduce cure by HSCT - viral rebound or death due to

underlying malignancy

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Elena Knops, AREVIR, Cologne 2017

The whole story (drama in 5 acts)

48y old patient, alive, well and undetectable for HIV (RNA/DNA) over 4 years after allogeneic CCR5-d32 HSCT

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Elena Knops, AREVIR, Cologne 2017

The beginning (prologue)

induction consolidation

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Elena Knops, AREVIR, Cologne 2017

The beginning (prologue)

induction consolidation

AML diagnosis in Jan 2011

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Elena Knops, AREVIR, Cologne 2017

The beginning (prologue)

induction consolidation

complete remission of AML

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Elena Knops, AREVIR, Cologne 2017

The beginning (prologue)

induction consolidation TDF/FTC/RAL to avoid interactions with chemotherapy

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Elena Knops, AREVIR, Cologne 2017

Chapter I

.... ....

  • ne week before

SCT

female donor, 10 out of 10 HLA- matched CCR5-d32

2 cycles high-dose cytarabine

September 2012

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Elena Knops, AREVIR, Cologne 2017

Chapter I

.... ....

  • ne week before

SCT

stem cells from female, 10/10 HLA-matched CCR5-d32 donor

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Elena Knops, AREVIR, Cologne 2017

Chapter I

.... ....

  • ne week before

SCT

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Elena Knops, AREVIR, Cologne 2017

Chapter I

.... ....

  • ne week before

SCT

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Elena Knops, AREVIR, Cologne 2017

Chapter II

.... during transplant and until today

  • n ART
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Elena Knops, AREVIR, Cologne 2017

Chapter II

....

total of 8 courses of 5-azacytidine + 4 donor lymphocyte infusions

June 2013

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Elena Knops, AREVIR, Cologne 2017

Chapter III

....

<LOD (884250) in rectal and ileum biopsy

10 10 10

+ CMV, HHV8, HSV

................ ................

2017

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Elena Knops, AREVIR, Cologne 2017

Chapter III

....

<LOD (884250) in rectal and ileum biopsy

10 10 10

+ CMV, HHV8, HSV

+ increased liver values ................

2017

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Elena Knops, AREVIR, Cologne 2017

Chapter III

....

<LOD (884250) in rectal and ileum biopsy

10 10 10

Tacrolimus + topical steroids + CMV, HHV8, HSV

................

2017

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Elena Knops, AREVIR, Cologne 2017

Chapter III

....

<LOD (884250) in rectal and ileum biopsy

+ CMV, HHV8, HSV

................

2017

Tacrolimus + topical steroids

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Elena Knops, AREVIR, Cologne 2017

Chapter III

....

<LOD (884250) in rectal and ileum biopsy

+ CMV, HHV8, HSV Tacrolimus + topical steroids

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Elena Knops, AREVIR, Cologne 2017

Chapter III

....

+ CMV, HHV8, HSV

  • PBMCs
  • rectal + ileum
  • bone marrow
  • CSF

Tacrolimus + topical steroids

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Elena Knops, AREVIR, Cologne 2017

Chapter III

....

2016 2017

ABC/3TC/DTG <20 <20 505 318

<LOQ (10 Mio) <LOQ (10 Mio) 1/10 repl <LOQ (3,8 Mio) <LOQ (15 Mio) 1/20 repl

52 month after SCT

<LOD (884250) in rectal and ileum biopsy

Tacrolimus + topical steroids

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Elena Knops, AREVIR, Cologne 2017

Chapter V

2016

Mar Apr May Jun

ICISTEM (M. Nijhuis, Utrecht): HIV LTR-DNA not detected in >2 Mio PBMCs via ddPCR or qPCR, but positive signals in T-cell subsets: TCM: 0.2 Mio cells (ddPCR) – 6.7cop/106cells TEM: 0.36 Mio cells (qPCR) – 5 cop/106cells

39

month after SCT

40 41 42

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Elena Knops, AREVIR, Cologne 2017

2016

  • G. Dunay, Hamburg:

Ileum biopsy: 0.1 Mio cells measured: 1/4 replicates positive with LTR-, but negative with gag primers ICISTEM (M. Nijhuis, Utrecht): HIV LTR-DNA not detected in >2 Mio PBMCs via ddPCR or qPCR, but positive signals in T-cell subsets: TCM: 0.2 Mio cells (ddPCR) – 6.7cop/106cells TEM: 0.36 Mio cells (qPCR) – 5 cop/106cells

Chapter V

Mar Apr May Jun

39

month after SCT

40 41 42

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Elena Knops, AREVIR, Cologne 2017

2016

ICISTEM (J. Martinez-Picado & M. Salgado, Barcelona): qVOA: 23 Mio CD4+ T cells negative (IUPM < 0.031/106 CD4 T cells) Ileum biopsy: 0.007 Mio CD45+ cells negative (<143/106 CD45+ cells) Rectum biopsy: 0.0297 Mio CD45+ cells negative (<34/106 CD45+ cells) ICISTEM (M. Nijhuis, Utrecht): HIV LTR-DNA not detected in >2 Mio PBMCs via ddPCR or qPCR, but positive signals in T-cell subsets: TCM: 0.2 Mio cells (ddPCR) – 6.7cop/106cells TEM: 0.36 Mio cells (qPCR) – 5 cop/106cells

  • G. Dunay, Hamburg:

Ileum biopsy: 0.1 Mio cells measured: 1/4 replicates positive with LTR-, but negative with gag primers

Chapter V

Mar Apr May Jun

39

month after SCT

40 41 42

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Elena Knops, AREVIR, Cologne 2017

2016

ICISTEM (J. Martinez-Picado & M. Salgado, Barcelona): qVOA: 23 Mio CD4+ T cells negative (IUPM < 0.031/106 CD4 T cells) Ileum biopsy: 0.007 Mio CD45+ cells negative (<143/106 CD45+ cells) Rectum biopsy: 0.0297 Mio CD45+ cells negative (<34/106 CD45+ cells) ICISTEM (M. Nijhuis, Utrecht): HIV LTR-DNA not detected in >2 Mio PBMCs via ddPCR or qPCR, but positive signals in T-cell subsets: TCM: 0.2 Mio cells (ddPCR) – 6.7cop/106cells TEM: 0.36 Mio cells (qPCR) – 5 cop/106cells

  • I. Hauber, Hamburg:

mVOA: negative in plasma, negative in histology

  • G. Dunay, Hamburg:

Ileum biopsy: 0.1 Mio cells measured: 1/4 replicates positive with LTR-, but negative with gag primers

Chapter V

Mar Apr May Jun

39

month after SCT

40 41 42

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Elena Knops, AREVIR, Cologne 2017

Rag#1DD Spleen Liver LN hu-CD3 HIVp24Ag

  • I. Hauber, Hamburg:

mVOA: negative in plasma, negative in histology

Chapter V

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Elena Knops, AREVIR, Cologne 2017

# sequencing genotypic prediction phenotypic analysis in Tcells Clone gp120-V3 amino acid sequence reads FPR (%) Magi (R5/X4) MT2 (X4) D1 CTRPNNNTREGIHIGPGRAFFTTGEIIGNIREASC 4 95,78 R5 R5 D2 CTRPNNNTRKSIHIGPGRAFFTTGEIIGNIKEAYC 2 95,64 R5 R5 D3 CTRPNNNTRKSIHIGPGRAFFTTGEIIGNIGEAYC 2 95,64 R5 R5 D4 CTRPNNNTRKGIHIGPGRAFFTTGEIIGNIREASC 2062 77,33 R5 R5 D5 CTRPNNNTRKGITIGPGRAFFTTGEIIGDIRQAHC 4886 30,67 R5 R5 D6 CTRPNNNTRKGIHIGSRKAFFTTGGIIGDIRQAYC 2 10,61 R5/X4 X4 D7 CTRPHTNTRKRIHIGPGRAFFTTGEIIGDIRQAYC 7 1,74 R5/X4 X4 D8 CTKPNNNTRKRIHIGPGRAFFTTGEIIGNIRQASC 2 1,74 R5 R5 D9 CTRPNNNIRKRIHIGPGRAFFTTGEIIGNIREAYC 3 1,16 R5/X4 X4

HxB2V3Bal

control R5 51,8 R5 R5 HxB2 control X4 X4 X4

Monique Nijhuis, Utrecht, ICISTEM PHENOTYPIC CO-RECEPTOR TROPISM ANALYSIS

(sample before transplantation, PBMC)

Chapter V

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Elena Knops, AREVIR, Cologne 2017

# sequencing genotypic prediction phenotypic analysis in Tcells Clone gp120-V3 amino acid sequence reads FPR (%) Magi (R5/X4) MT2 (X4) D1 CTRPNNNTREGIHIGPGRAFFTTGEIIGNIREASC 4 95,78 R5 R5 D2 CTRPNNNTRKSIHIGPGRAFFTTGEIIGNIKEAYC 2 95,64 R5 R5 D3 CTRPNNNTRKSIHIGPGRAFFTTGEIIGNIGEAYC 2 95,64 R5 R5 D4 CTRPNNNTRKGIHIGPGRAFFTTGEIIGNIREASC 2062 77,33 R5 R5 D5 CTRPNNNTRKGITIGPGRAFFTTGEIIGDIRQAHC 4886 30,67 R5 R5 D6 CTRPNNNTRKGIHIGSRKAFFTTGGIIGDIRQAYC 2 10,61 R5/X4 X4 D7 CTRPHTNTRKRIHIGPGRAFFTTGEIIGDIRQAYC 7 1,74 R5/X4 X4 D8 CTKPNNNTRKRIHIGPGRAFFTTGEIIGNIRQASC 2 1,74 R5 R5 D9 CTRPNNNIRKRIHIGPGRAFFTTGEIIGNIREAYC 3 1,16 R5/X4 X4

HxB2V3Bal

control R5 51,8 R5 R5 HxB2 control X4 X4 X4

Monique Nijhuis, Utrecht, ICISTEM CO-RECEPTOR TROPISM

(sample before transplantation, PBMC)

Chapter V

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Elena Knops, AREVIR, Cologne 2017

The end? (epilogue)

  • qVOA + ultra microchimerism (ICISTEM: M. Salgado, Barcelona):

lymph node (planned)

  • T-cell response assay (T. Harrer, Erlangen):

PBMCs from donor (in progress)

  • proviral DNA load from further PBMC samples and lymph node

(Institute of Virology, Cologne and others)

  • …

International collaboration to guide and investigate the potential for HIV cure by stem cell transplantation. The ICISTEM project is supported by amfAR Research Consortium on HIV eradication (ARCHE) Research Grant # 108930-56-RGRL www.icistem.org

More assays to be done:

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Elena Knops, AREVIR, Cologne 2017

Institute for Immunogenetics, Kaiserslautern

Martin Däumer, Alex Thielen

Heinrich Pette Institut HH

Ilona Hauber, Gabor Dunay, Johanna Eberhard, Silke Kummer, Julian Schulze zur Wiesch

Med 3, University of Erlangen

Thomas Harrer

IciStem project

Annemarie Wensing, Javier Martinez-Picado, Monique Nijhuis et al.

MPI of Informatics, Saarbrücken

Thomas Lengauer, Joachim Büch, Nico Pfeifer, Alejandro Pironti, Prabhav Kalaghatgi

Thanks to our group

Institute of Virology, University of Cologne

Rolf Kaiser Eva Heger Veronica Di Cristanziano Maria Neumann-Fraune Eugen Schülter Claudia Müller Saleta Sierra-Aragon Sascha Hasheminasab

University of Düsseldorf

Hepatology & Infectiology Björn Jensen Falk Hüttig Dieter Häussinger Virology Nadine Lübke Haematology Guido Kobbe Moleculare Medizin Philipp Lang Junnat Hamdam

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Elena Knops, AREVIR, Cologne 2017

Thanks to the „Duesseldorf“ patient

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Elena Knops, AREVIR, Cologne 2017

02/2013

  • pos. control
  • neg. control

06/2014 02/2015 07/2015 12/2015 01/2016

gp160 gp120 gp41 p34 p25 p18 p55 p68

Intermezzo

before SCT